Rapid and effective separation of nucleotides (NTs) and their derivatives is crucial for studying their physiological functions. In this work, we comprehensively evaluated the separation ability of a zwitterionic hydrophilic monolith, i.e., poly(N,N-dimethyl-N-(3-methacrylamidopropyl)-N-(3-sulfopropyl)ammonium betaine-co-N,N'-methylenebisacrylamide) (poly(SPP-co-MBA)) for NTs analysis, including its selectivity, chemical stability under extremely basic condition and compatibility with hydrophilic interaction liquid chromatography (HILIC) coupled with mass spectrometry (HILIC-MS). The poly(SPP-co-MBA) monolith exhibited excellent chemical stability, as evidenced by the low relative standard deviation of retention time (0.16-1.05%) after 4000 consecutive injections over one month under strong alkaline elution condition (pH 10). After optimizing the separation conditions, including buffer pH and concentration, organic solvent content and column temperature, four nucleoside triphosphates, five nucleoside diphosphates and five nucleoside monophosphates were baseline separated within 7 min. Additionally, the mixtures containing one nucleoside and its corresponding mono-, di-, and triphosphates were baseline separated within only 3 min, respectively. It is good HILIC-MS compatibility was also confirmed by the satisfactory peak shape and high response of nine NTs. Overall, the proposed poly(SPP-co-MBA) monolith exhibited good mechanical stability and compatibility of HILIC-MS, making it a promising technique for NTs analysis.

中文翻译：

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用于核苷酸分离的聚合两性离子亲水整体料的综合评价。

快速有效地分离核苷酸（NT）及其衍生物对于研究其生理功能至关重要。在这项工作中，我们综合评价了两性离子亲水性整体材料，即聚(N,N-二甲基-N-(3-甲基丙烯酰胺丙基)-N-(3-磺基丙基)铵甜菜碱-co-N,N'的分离能力。 -亚甲基双丙烯酰胺）（聚（SPP-co-MBA））用于 NT 分析，包括其选择性、极碱性条件下的化学稳定性以及与亲水相互作用液相色谱 (HILIC) 联用质谱 (HILIC-MS) 的兼容性。聚（SPP-co-MBA）整体材料表现出优异的化学稳定性，强碱性洗脱条件（pH 10）下一个月内连续进样 4000 次后保留时间的相对标准偏差较低（0.16-1.05%）证明了这一点。优化分离条件（包括缓冲液 pH 和浓度、有机溶剂含量和柱温）后，四种核苷三磷酸、五种核苷二磷酸和五种核苷单磷酸在 7 分钟内实现基线分离。此外，含有一个核苷及其相应的一磷酸、二磷酸和三磷酸的混合物分别在 3 分钟内即可实现基线分离。9 个 NT 的令人满意的峰形和高响应也证实了其良好的 HILIC-MS 兼容性。总体而言，所提出的聚（SPP-co-MBA）整体材料表现出良好的机械稳定性和 HILIC-MS 兼容性，使其成为 NT 分析的一种有前途的技术。